The permeability of the podocyte monolayer to albumin was measured as previously described (94 (link)). Briefly, podocytes were seeded in the upper chambers of 3-μm polycarbonate Transwell filters of a 24-well filtration microplate (Corning). Four days after 4OHT treatment, cells were washed with PBS supplemented with 1 mM MgCl2 and 1 mM CaCl2 to protect the cadherin-based junctions. The top chamber was filled with 0.15 ml of complete medium, and the bottom chamber was filled with 750 μl of complete medium supplemented with bovine serum albumin (BSA; 40 mg/ml). Two hours later, the medium from the upper chamber was collected, and the albumin concentration was measured using the Pierce Rapid Gold BCA Protein Assay Kit (Thermo Fisher Scientific, A53225).
24 well filtration microplate
Manufactured by Corning
The 24-well filtration microplate is a laboratory equipment designed for efficient sample processing. It features 24 individual wells, each equipped with a filter membrane, enabling the separation of solid and liquid components in multiple samples simultaneously. The core function of this product is to facilitate the filtration and purification of various substances, supporting researchers and scientists in their analytical and experimental workflows.
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2 protocols using 24 well filtration microplate
1
Podocyte Permeability to Albumin
2
Measuring Podocyte Monolayer Permeability to Albumin
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The permeability of podocyte monolayer to albumin was measured as previously described [4] (link), [49] (link). Briefly, podocytes were seeded in the upper chambers of 3 µm polycarbonate Transwell filters of a 24-well filtration microplate (Corning, New York, NY). After HG treatment with or without PRR siRNA for 72 h, cells were washed with PBS supplemented with 1 mmol/L MgCl2 and 1 mmol/L CaCl2 to protect the cadherin-based junctions. The top chamber was filled with 0.15 ml of fresh RPMI 1640 and the bottom chamber with 1 ml of RPMI 1640 supplemented with 40 mg/ml of bovine serum albumin. 2 hours later, the medium from the upper chamber was collected, and the albumin concentration was measured using Bio-Rad protein assay (Bio-Rad).
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